NOVEL MODULATORS OF THE VANILLOID RECEPTOR

香草酸受体的新型调节剂

• 批准号: 7083038
• 负责人: Joseph C Glorioso
• 金额: $ 41.34万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7083038
• 关键词: Alphaherpesvirinae; analgesia; apoptosis; behavior test; calcium; capsaicin; complementary DNA; diabetic neuropathy; dorsal root; gene therapy; genetic library; genetic screening; genetic transduction; laboratory rat; membrane channels; pain; plasmids; spinal ganglion; transfection /expression vector; urinary tract; western blottings

The vanilloid receptor (TRPV1; formerly known as VR1) plays an important role in primary hyperalgesia, a component of chronic pain. TRPV1 is functionally up-regulated in patho-physiologic states such as painful diabetic neuropathy and cystopathy. Activation of TRPV1 occurs through a number of convergent signaling pathways and emerging evidence suggests that TRPV1 function is also negatively regulated by numerous mechanisms. The discovery of novel cellular inhibitors or negative modulators of TRPV1 function and their mode of action should provide additional insights into the role of TRPV1 in peripheral pain signaling and by extension, suggest novel approaches for their application to the control of primary hyperalgesia. We propose to test the hypotheses that (i) novel cellular products play an important role in the inhibition or negative modulation of TRPV1 function and (ii) that vector-based expression of these molecules can control TRPV1 signaling in vivo. To test these hypotheses, we have devised a combination of HSV vector-based genetic strategies to identify these potential regulatory molecules and biochemical methods to characterize their role in the negative control of TRPV1 function. Specifically, we will attempt to identify products that (a) inhibit TRPV1 activation by capsaicin (CAP) and resiniferatoxin (RTX) and (b) interfere with TRPV1 potentiation by protein kinase C epsilon (PKCe) activated by Phorbol 12-myristate 13-acetate (PMA). TRPV1 inhibitory genes that either have been engineered or naturally occur, will be studied in some detail to determine the molecular mechanism underlying their ability to impede TRPV1-mediated calcium influx. In four specific aims we intend to (i) create model systems in which HSV TRPV1 expression vectors can be used to examine potential inhibitors of TRPV1 function or calcium overload (ii) create a library of vectors expressing cDNAs derived from rat dorsal root ganglia, (iii) characterize mechanisms by which the selected neuronal gene products inhibit or negatively modulate TRPV1 function and (iv) evaluate the analgesic effects of the engineered and novel TRPV1 inhibitors in vivo using rat models of pain where TRPV1 antagonism has been shown to reduce pain signaling. Both the model inhibitor genes and novel genes obtained by the HSV screening methods will be provided to Projects 1 and 2 for evaluation of their potential analgesic effects in diabetes-related neuropathic pain and models of bladder pain.

香草酸受体（TRPV1；以前称为 VR1）在原发性痛觉过敏中发挥重要作用，原发性痛觉过敏是一种 慢性疼痛的组成部分。 TRPV1 在病理生理状态（例如疼痛）中功能上调 糖尿病神经病变和膀胱病。 TRPV1 的激活是通过许多汇聚信号传导发生的 途径和新出现的证据表明 TRPV1 功能也受到多种因素的负调控 机制。 TRPV1 功能的新型细胞抑制剂或负调节剂的发现及其作用 作用模式应该提供更多关于 TRPV1 在外周疼痛信号传导中的作用的见解，并通过 扩展，提出了用于控制原发性痛觉过敏的新方法。我们建议 检验以下假设：(i) 新型细胞产物在抑制或负面作用中发挥重要作用 TRPV1 功能的调节以及 (ii) 这些分子的基于载体的表达可以控制 TRPV1 体内信号传导。为了检验这些假设，我们设计了基于 HSV 载体的遗传组合 识别这些潜在调控分子的策略和表征其作用的生化方法 TRPV1 功能的负控制。具体来说，我们将尝试识别能够 (a) 抑制的产品 辣椒素 (CAP) 和树脂毒素 (RTX) 激活 TRPV1，并且 (b) 通过以下方式干扰 TRPV1 增强： 蛋白激酶 C epsilon (PKCe) 由佛波醇 12-肉豆蔻酸酯 13-乙酸酯 (PMA) 激活。 TRPV1 抑制 基因工程或自然发生的基因，将进行一些详细的研究，以确定 它们阻碍 TRPV1 介导的钙内流能力的分子机制。四个具体目标 我们打算 (i) 创建模型系统，其中 HSV TRPV1 表达载体可用于检查 TRPV1 功能或钙超载的潜在抑制剂 (ii) 创建表达 cDNA 的载体文库 源自大鼠背根神经节，（iii）表征所选神经元基因的机制 产品抑制或负向调节 TRPV1 功能，并 (iv) 评估产品的镇痛效果 使用大鼠疼痛模型在体内设计新型 TRPV1 抑制剂，其中 TRPV1 拮抗作用已被证实 显示可以减少疼痛信号。 HSV获得的模型抑制基因和新基因 将向项目1和2提供筛选方法，以评估其潜在镇痛效果 糖尿病相关的神经性疼痛和膀胱疼痛模型。